ABSTRACT
The study was undertaken to evaluate milk clotting and proteolytic activity of protein fractions of seeds of Lagenaria siceraria. Protein fractions were isolated by the method of differential solubility, and their protein contents were estimated using Bio-Rad protein assay reagent and bovine serum albumin. The effects of pH and temperature on the milk-clotting and proteolytic activity were also evaluated. The isolated protein fractions showed highest milk clotting activity over a broad temperature range of 30-80 ºC and pH range of 3-9. Based upon the observations, milk clotting enzymes present in different fraction of proteins of Lagenaria siceraria are promising candidates for application in industrial scale for production of cheese and might be a potential substitute for commercial animal rennet.
ABSTRACT
Breadfruit is an exotic tree in Brazil, very well acclimatized. Despite of being a laticifer plant, the knowledge about its latex is scarce. However, it is known that proteolytic enzymes represents over 50% of the latex composition in laticifers plants. The aim of this study was to evaluate the potential of breadfruit (Artocarpus altilis var. Apyrena) latex as a source of milk clotting proteases and partially characterize it. The proteolytic activity of the fraction of crude extract was assessed using azocasein and quantitation of total protein, the bicinchoninic acid (BCA). Milk-clotting activity was analyzed using skim milk at 12%. The enzyme activity was analyzed under different temperature conditions (35 to 80°C) and pH (5.8 to 10.7) presenting optimum activity in alkaline pH (8.5) and 50°C; being stable to the two variables at 120 minutes during the test. The clotting activity was directly proportional to the temperature at the better concentration of CaCl2 (10mmol L-1). The results indicate that enzyme is a possible replacement for calf rennet.
A fruta-pão é uma árvore exótica no Brasil, onde se aclimatou muito bem. Embora seja uma planta lactífera, o conhecimento sobre seu látex é escasso. No entanto, sabe-se que enzimas proteolíticas correspondem a mais de 50% da composição do látex em plantas lactíferas. O objetivo deste estudo foi avaliar o potencial do látex da fruta-pão (Artocarpus altilis var. Apyrena) como fonte de protease coagulante do leite e caracterizar parcialmente a enzima. A atividade proteolítica da fração de extrato bruto foi avaliada utilizando azocaseína e, para a quantificação das proteínas totais, o ácido bicinconínico (BCA). A atividade de coagulação do leite foi testada utilizando leite desnatado a 12%. A protease foi testada em diferentes condições de temperatura (35 a 80°C) e pH (5,8 a 10,7) e apresentou atividade ótima a 50°C e pH alcalino (8,5) sendo estável a estas variáveis durante 120 minutos. A atividade coagulante no leite foi diretamente proporcional à temperatura na melhor concentração de CaCl2 a 10µmol L-1. Os resultados indicam que a enzima analisada é uma possível alternativa à quimosina.
ABSTRACT
Antecedentes: en Colombia, el alto recuento de células somáticas (RCS) en la leche es un problema para la industria lechera. Altos recuentos pueden afectar de manera considerable los rendimientos y calidad final del queso. Varios países han establecido límites máximos estandarizados para el RCS. Colombia no lo ha hecho de manera oficial y tan solo unas pocas industrias manejan sus propios límites. Objetivos: Determinar el efecto del RCS sobre parámetros de aptitud quesera de la leche y la calidad sensorial del queso campesino. Métodos: Se tomaron muestras de leche de seis tanques con altos y bajos RCS y se realizaron mezclas para obtener 30 baches con diferentes RCS (desde 150.000 hasta 1.200.000 cel/ml). Con estas mezclas se elaboraron 30 quesos tipo campesino a los cuales se les determinaron variables de aptitud quesera (tiempo de coagulación, rendimientos y pérdidas en suero) y la calidad organoléptica a través de una prueba sensorial descriptiva de puntajes con panel de seis jueces con experiencia previa y entrenados en queso campesino. Las variables de aptitud quesera fueron analizadas por regresión múltiple y los resultados de la evaluación sensorial con la prueba no paramétrica de Friedman. Resultados: La aptitud quesera disminuyó con RCS superiores a 200.000 cel/ml. El tiempo de coagulación (R² = 0.74; P< 0.001) y las pérdidas de proteína en el lactosuero (R² = 0.55; P<0,001) aumentaron, mientras que los rendimientos (R²=0.31; P<0.01) disminuyeron a medida que aumentó el RCS. La calificación de los panelistas respecto de la textura y la apariencia disminuyó con RCS mayores a 600.000 cel/ml (P<0.01) y el sabor y el aroma, a partir de 800.000 cel/ml (P<0,01). Conclusiones: Aumentos en el RCS en leche afectan negativamente parámetros de aptitud quesera y la calidad sensorial del queso campesino. Se sugiere que los impactos serán menores sobre el rendimiento cuanto menor sea el RCS, mientras que la calidad organoléptica mejorará cuando la leche tenga RCS por debajo de 600.000 cel/ml.
Background: A high milk somatic cell count (SCC) is a problem for milk industry in Colombia. These high counts can affect considerably yield and final quality of cheese. Several countries have established maximum limits for SCC. In Colombia these limits have not been established officially, only a few industries have their own limits. Objectives: To determine the effect of somatic cell count (SCC) on milk potential for cheese making and sensorial quality of fresh cheese. Methods: Six milk samples with high and low SCC, were taken from bulk tanks and mixed to obtain 30 samples with SCC of 150.000 to 1.200.000 somatic cells/mL. Thirty fresh cheeses were prepared and clotting time, yield (protein, fat, dry matter) and whey losses were determined. Additionally, score descriptive sensorial quality test was performed by 6 trained judges. Protein, fat, dry matter in milk and cheese yields were analyzed by multiple regressions and information of sensorial test by Friedman method. Results: When milk SCC (somatic cells/ml) increased from 150.000 to 1.200.000, clotting time (R² = 0,74; P<0.001), and whey protein losses increased (R² = 0.55; P<0.001) and cheese yield decreased (R²=0.31, P<0.01). According to panelists, texture and appearance were affected negatively when SCC was higher than 600.000 cells per ml (P<0.01), flavor and aroma when they were higher than 800000 cells/ml. Conclusions: Increases in SCC have a negative effect on milk potential for cheese making and quality sensorial parameters on fresh cheese. It is suggested that minor impacts in fresh cheese yield would be obtained with a lower SCC and for good sensorial quality when the milk has SCC, lower than 600.000 cells per ml.
ABSTRACT
Objective To evaluate the efficacy and safety of composite milk clotting enzyme capsules in children with functional dyspepsia.Methods A multicenter,randomized,open-label clinical trial was carried out in 4Guangzhou hospitals during Feb.2012 to Mar.2013.Children with functional dyspepsia on basis of Rome Ⅲ criteria were enrolled and treated with composite milk clotting enzyme capsules.Total symptom score and drug-related adverse reactions were evaluated after one-week and two-week therapy.The treatment outcome were divided into very effective,fairly effective and ineffective according to the symptom score changes.Superiority test was done for effectiveness of the treatment based on efficacy difference by 10％ regarded as superiority.Results Two hundred and one children were enrolled.One hundred and ninety-six children completed the study as planned.After two-week therapy,better effective rate and total effective rate were 68.88％ and 87.76％,higher than those of one-week therapy (27.04％,76.02％) (U =2.935,P ＜ 0.05).The lower limits of 95 ％ confidence interval of difference in symptoms improve rate,better efficacy rate and symptom score decrease rate between two-week and one-week therapy were more than 10％,and two-week therapy was superior to one-week therapy.No significant drug-related adverse reaction was found during the study.Conclusions Composite milk clotting enzyme capsules can improve functional dyspepsia symptoms significantly.The effectiveness of two-week of treatment is better than that of one-week treatment.No obvious drug-related adverse reaction is observed.
ABSTRACT
The partial characterization and purification of milk clotting enzyme obtained from the (root latex) of Jacaratia corumbensis O. kuntze was studied, by fractional precipitation with ammonium sulphate and ion exchange chromatography. The ammonium sulphate precipitate showed five fractions (AS1- 0-20 percent; AS2 - 20-40 percent; AS3 - 40-60 percent; AS4 - 60-80 percent; AS5 - 80-100 percent) and among the fractions obtained, the 40-60 percent fraction (AS3) showed the highest milk clotting activity with a purification factor of 1.2 fold in relation to the crude extract. This fraction when applied on Mono Q column yielded two protein peaks (p1 and p2), but p1 pool showed the best milk-clotting activity. The optimal pH for the crude and partially purified extract was 6.5 and 7.0, respectively. The maximum milk-clotting activity was at 55ºC for the both crude and partially purified extracts. The enzyme was inhibited by iodoacetic acid which suggested that this enzyme was a cysteine protease, with molecular weight of 33 kDa.
A enzima coagulante de leite obtida de látex de raiz de Jacaratia corumbensis O. kuntze foi caracterizada parcialmente e purificada, por precipitação fracionária com sulfato de amônio e cromatografia de troca de íon. Foram utilizadas cinco frações de sulfato de amônio (AS1 - 0-20 por cento; AS2 - 20-40 por cento; AS3 - 40-60 por cento; AS4 - 60-80 por cento; AS5 - 80-100 por cento), a fração 40-60 por cento (AS3) mostrou alta atividade coagulante com um fator de purificação de 1,2 vezes em relação ao extrato bruto. Esta fração foi aplicada em coluna Mono Q obtendo dois picos de proteína (p1 e p2), o p1 mostrou melhor atividade coagulante. O pH ótimo para o extrato bruto e parcialmente purificado foi 6,5 e 7,0, respectivamente. A atividade coagulante foi atingida a 55ºC para ambos os extratos, bruto e parcialmente purificado. A enzima foi inibida por ácido iodoacético que sugere que esta enzima é uma cisteína protease, com peso molecular de 33 kDa.